WO1991009046A1

WO1991009046A1 - Morpholinyl derivatives of doxorubicin and process for their preparation

Info

Publication number: WO1991009046A1
Application number: PCT/EP1990/002229
Authority: WO
Inventors: Alberto Bargiotti; Maria Grandi; Antonino Suarato; Daniela Faiardi
Original assignee: Farmitalia Carlo Erba SRL; Carlo Erba SpA
Current assignee: Pfizer Italia SRL
Priority date: 1989-12-19
Filing date: 1990-12-18
Publication date: 1991-06-27
Anticipated expiration: 1992-06-19
Also published as: SK621990A3; IL96643A0; IL111527A; NO176911C; ES2094136T3; CN1079215A; ATE143023T1; FI97389B; HRP921226B1; NO913176D0; NO175533C; IL111527A0; DE69028610D1; PT96226B; CN1039708C; KR0183033B1; NO913176L; TW208019B; YU237990A; HUT59693A

Abstract

Morpholinyl derivatives of doxorubicin having formula (A), in which X represents a linear or branched C2?-C6? alkyl group or a benzyl group -CH2?C6?H5? and which have (S) or (R) configuration at carbon atom C-2'' of the morpholino ring, are antitumor agents.

Description

MORPHOLINYL DERIVATIVES OF DOXORUBICIN AND PROCESS FOR THEIR

PREPARATION

The invention relates to anthracycline glycosides, to processes for their preparation and to pharmaceutical compositions containing them.

The invention provides new anthracycline glycosides of general formula A in which the 3'-nitrogen atom is enclosed in a 2-alkoxy-4-morpholinyl ring:

in which X represents a linear or branched C₁-C₆ alkyl group or a benzyl residue -CH₂C₅H₅ and which has a (S) or (R) configuration at the 2"-carbon atom, and pharmaceutically acceptable acid addition salts thereof. The preferred salt is the hydrochloride salt.

Morpholino-anthracyclines are well known compounds endowed with promising antitumor activity on experimental murine tumors [see: E.W. Acton in Bioactive Molecules, 55- 101, vol 6, Edited by J.W. Lown, Elveiser 1988]. Among these, 2-methoxy-4-morpholinyl anthracyclines (X = OCH₃) have been already claimed in our patent US-A-4,672,057.

These compounds were prepared through a reductive-alkylating process using a chiral-dihaldehyde. In the present

invention, on the other hand, the substituted morpholinyl rings are prepared through bis-alkylation of the 3'-amino group of anthracyclines with novel chiral 1,5-diiodo-2- alkoxy or -benzyloxy derivatives that are within the scope of the invention.

The preferred anthracycline glycosides of general formula A include:

A1: 3'-deamino-3'-(2"(S)-benzyloxy-4"-morpholinyl)- doxorubicin (X=CH₂C₆H₅),

A2 : 3'-deamino-3'-(2"(S)-ethoxy-4"-morpholinyl)-doxorubicin (X=C₂H₅),

A3 : 3'-deamino-3'-(2"(R)-isopropyloxy-4"-morpholinyl)- doxorubicin (X=CH(CH₃)₂),

A4: 3'-deamino-3'-(2"(S)-methoxy-4"-morpholinyl)- doxorubicin (X=CH₃), and

A5: 3'-deamino-3'-(2"(R)-methoxy-4"-morpholinyl)- doxorubicin (X=CH₃).

and their hydrochloride salts. The compounds may have a (S) or (R) configuration at carbon atom C-2" of the morpholino ring.

The new anthracycline glycoside antibiotics of the invention, i.e. those of general formula A., are prepared by the formation of a substituted morpholinyl ring at C-3' on the sugar moiety of the antitumor anthracycline glycoside doxorubicin (B) :

The present invention therefore provides a process for the preparation of an anthracycline glycoside of formula A or a pharmaceutically acceptable acid addition salt thereof, which process comprises:

(i) reacting doxorubicin or an acid addition salt thereof, for example the hydrochloride salt, with a diiodo compound of general formula C:

wherein X is as defined above; and

(ii) if desired, converting the anthracycline glycoside of formula A thus obtained into a pharmaceutically acceptable acid addition salt thereof.

The alkylation of the C-3' amino group of doxorubicin or the doxorubicin salt is typically performed in step (i) in a polar aprotic solvent and in the presence of a dry organic base such as triethylamine. Reaction is generally carried out at room temperature from eight to twenty four hours. The carbon atom C-2 bearing the -OX group in the diiodo compound may have a (S) or (R)

configuration. In a preferred embodiment, doxorubicin or its hydrochloride, dissolved in a polar aprotic solvent is reacted, at room temperature and in the presence of a dry organic base, with the diiodo compound of general formula C to give the corresponding morpholinyl doxorubicin derivative of formula A which, after purification on a silica gel column using as eluting system methylene chloride-methanol (97:5 v/v), is isolated, by treatment with methanolic anhydrous hydrogen chloride, as its hydrochloride. A pure 2"(R)-[(C₁-C₆) alkoxy or benzyloxy]-anthracycline glycoside of formula A or salt thereof, or a pure 2"(S)-[(C₁-C₆) alkoxy or benzyloxy]-anthracycline glycoside of formula A or salt thereof, may therefore be provided according to the present invention.

The invention also provides a process for the preparation of optically pure diiodo compounds C, starting from sugar precursors such as the compound of general formula S derived from L-arabinose:

wherein X is as defined above. This process comprises:

(a) subjecting to periodate oxidation a compound of formula S¹:

wherein X is as defined above;

(b) reducing the thus-obtained dialdehyde derivative of formula D¹:

wherein X is as defined above;

(c) sulfonating the thus-obtained dihydroxy derivative of formula E¹:

wherein X is as defined above; and

(d) iodinating the sulfonated derivative thus obtained.

In order to prepare the diiodo compounds C, 1- substituted sugars S¹, prepared following standard

procedures described in "Methods on Carbohydrate Chemistry" Acad. Press., Vol 1, (1962), are first transformed into dialdehyde derivatives D¹. Generally, D- or L-arabinose is employed as a starting material. This is reacted with an alcohol X-OH thereby to form the compound of formula S¹.

The dialdehyde derivatives can be obtained by using

periodate oxidation in water, then reduced to 1,5-dihydroxy- 2-alkoxy or -benzyloxy-3-oxa-pentane E¹ by using reducing agents such as sodium borohydride or sodium cyanoborohydride at pH 6.5 in a mixture of water and methanol.

The resultant dihydro compounds E¹ are sulfonated at the 1- and 5-hydroxyl groups, typically by using ptoluensulfonyl chloride in pyridine at 4°C to give the sulfonyl esters of formula F from which the diiodo

derivatives C are obtained upon treatment with sodium or potassium iodide in aprotic solvent such as

methylethylketone at 85°C from one to two days. The

sequence of these reactions do not affect the chirality at

C-2 of the diiodo derivatives C which is the same of the starting sugars S.

A preferred embodiment of a process according to the invention is illustrated by the following reaction

Scheme:

The present invention also provides pharmaceutical compositions comprising a pharmaceutically acceptable carrier or diluent and, as active ingredient, an

anthracycline glycoside of formula A or a pharmaceutically acceptable acid addition salt thereof. Conventional

carriers or diluents may be used. The composition may be formulated and administered, for example intravenously, in conventional manner.

The anthracycline glycosides of formula A and pharmaceutically acceptable acid addition salts thereof are antitumour agents. They may be used to treat a patient with a tumour by administration of a therapeutically effective amount thereof. The compounds can be used to inhibit the growth of a tumour and are non-toxic at therapeutic doses.

The following Examples illustrate the invention.

Example 1

Preparation of 1,5-di(p-toluensulphonyl)oxy-2(s)-benzyloxy- 3-oxa-pentane (F1)

L-arabinose (3 g, 0.022 mole) and benzyl alcohol (15 ml) were heated under stirring in presence of Dowex 50Wx2 (2 g) in acidic form. After four hours, the mixture was cooled and filtered. The solvent was removed under reduced pressure and 1-benzyl-β-L-arabinopyranoside (S1, 3.5 g) was recovered from acetone. TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system methylene chloride/methanol/water (120/20/2 by volume) Rf=0.47. [α]_D=+215° (c=1% water).

1-benzyl-,9-L-arabinopyranoside (S1, 3.48 g, 0.0145 mole) was dissolved in water (100 ml) and treated with sodium periodate (5.6 g, 0.026 mole) at 0°C for two hours. Then barium chloride was added and the mixture was brought to pH 7 with barium carbonate, filtered off and washed with water. The aqueous solution was concentrated under reduced pressure to a syrup and extracted with acetonitrile (50 ml). The organic phase was diluted with a mixture of methanol (20 ml) and water (10 ml) and treated with sodium

cyanoborohydride (0.3 g) dissolved in water (5 ml). After 15 minutes the mixture was brought to pH 7 by adding Dowex 50Wx2 and filtered. The solvents were removed under reduced pressure to give 1, 5-dihydroxy-2(S)-benzyloxy-3-oxa-pentane (E1, 2.6g, yield 85%). TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system methylene chloride/methanol (10/1 by volume) Rur: Rf=0.28. Compound E1 (2.6 g) was dissolved in dry pyridine and added

with p-toluensulfonylchloride (6.67 g). The mixture was kept at O°C overnight, then poured into ice-water and extracted with methylene chloride. The organic phase was washed with water, separated off, dryied over anhydrous sodium sulphate and filtered off. The solvent was removed under reduced pressure to afford 1,5-di(p-toluensulρhonyl)oxy-2(S)-benzyloxy- 3-oxa-penta.ne (F1, 4.3 g, yield 68%). TLC on Kieselgel Plate F₂₉₄ (Merck), eluting system methylene chloride/acetone (98/2 by volume) R_f=0.55

1HNMR (CDCI₃, 400 MHz) δ:

2.42 (s, 6H, two CH₃Ts); 3.65, 3.69 (two dt, J=4.7, 11.7Hz, 2H, TsOCH₂CH₂O); 3.94 (two, dd, J=5.3, 10.5Hz, 2H, OCH-CH₂-OTs); 4.08 (t, J=4.7Hz, 2H, Ts-O-CH₂CH₂-O); 4.46, 4.56 (two, d, J=11.7Hz, 2H, OCH₂-Ph); 4.72 (t, J=5.3Hz, 1H, O-CH-CH₂-OTs); 7.2-7.8 (m, 13H, aromatics).

Example 2

Preparation of 1,5-diiodo-2(S)-benzyloxy-3-oxa-pentane (C1).

Compound F1 (4.3 g, 8.3 mmole), prepared as described in Example 1, was dissolved in methylethylketone (50 ml) and added with sodium iodide (7.4 g, 49 mmole). The mixture was kept at 95°C for twentyfour hours. After that, the solvent was removed under reduced pressure and the residue was extracted with n-hexane. The organic phase was concentrated to a syrup to give the title compound C1 (3.5 g, yield 90%). TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system n-hexane/ ethyl acetate (10/0.5 by volume) R_f=0.34

1HNMR (CDCI₃, 400 MHz) δ:

3.27 (t, J=6.8Hz, 2H, J-CH₂CH₃-O); 3.30 (d, J=5.5Hz, 2H, O-CH-CH_a-J); 3.84 (m, 2H, J-CH₂CH₂-O); 4.63, 4.74 (two d, J=11.7Hz, 2H, O-CH₂Ph); 4.81 (t, J=5.5Hz, 1H, O-CH-CH₂-J); 7.3-7.5 (m, 5H, aromatics). Example 3

Preparation of 3'-deamino-3'[2(S)-benzyloxy-4-morpholinyl] doxorubicin (A1)

To a solution of doxorubicin hydrochloride (0.5 g, 0.86 mmole) in dry dimethylformamide (20 ml) was added 1,5-diiodo- -2(S)-benzyloxy-3-oxa-pentane (cl, 3.5 g, 7.54 mmole) and dry triethylamine (3.6 ml, 2.6 mmole). The mixture was kept at room temperature for 36 hours, then was poured in water and extracted with methylene chloride. After standard work-up, the crude product was purified on silicic acid column using as eluting system a mixture of methylene chloride/methanol (97/5 by volume), to give, after treatment with methanolic anhydrous hydrogen chloride, the title compound A1 (0.3 g, yield 46%) as hydrochloride salt.

TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system methylene chloride/methanol (10/1 by volume) R_f=0.6.

FD-MS: m/e 756 (M+)

¹HNMR of free base (CDCl₃, 200 MHz) δ:

1.37 (d, J=6.6Hz, 3H, 5'-CH₃); 1.76 (m, 2H, 2'-CH₂); 2.14 (dd, J=3.9, 14.8Hz, 1H, 8ax-H); 2.2-2.7 (m, 6H, CH₂-N-CH₂, 8eg-H, 3'-H); 3.00 (d, J=18.8Hz, 1H, 10ax-H); 3.55, 4.00 (two m, 2H, O-CH₂CH₂N); 3.68 (s, 1H, 4'-H); 3.94 (g, J=6.6Hz, 1H, 5'-H); 4.08 (s, 3H, OCH₃); 4.51, 4.77( two d, J=12.1Hz, 2H, OCH₂Ph); 4.65 (dd, J=2.6, 4.0Hz, 1H, OCH(OCH₂Ph)CH₂N); 4.71 (s, 1H, COCH₂OH); 5.28 (dd, J=2.4, 3.8Hz, 1H, 7-H); 5.54 (m, 1H, 1'-H); 7.2-8.1 (m, 8H, aromatic H's); 13.22 (s, 1H, 11-OH); 13.95 (s, 1H, 6-OH). Example 4

Preparation of 1,5-diiodo-2(S)-ethoxy-3-oxa-pentane (C2).

The title compound C2 was prepared starting from L-arabinose (3 g) following sequential reactions as described in Example 1 and Example 2.

1-ethyl-α-L-arabinopyranoside (S2) : [α]_D=+233.5° (c=1%water) 1,5-dihydroxy-2(S)-ethoxy-3-oxa-pentane (E2): TLC on KieselgelPlate F₂₅₄ (Merck) eluting system methylene chloride/methanol (10/1 by volume) R_f=0.33.

1,5-di(p-toluensulphonyl)oxy-2(S)-ethoxy-3-oxa-pentane (F2): TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system methylene chloride/acetone (98/2 by volume) R_f=0.56

1HNMR (CDCl₃, 200MHz) δ:

1.11 (t, J=7.0HZ, 3H, OCH₂CH₃); 2.43 (s, 6H, two CH₃-Ts); 3.44, 3.58 (two dg, J=7.0, 9.4Hz, 2H, OCH₂CH₃); 3.68 (m, 2H, O-CH₂-CH₂-OTs); 3.89 (d, J=5.4, 2H, o-CH-CH₂-OTs); 4.10 (t, J=4.8Hz, 2H, OCH₂-CH₂OTs); 4.61 (t, J=5.4Hz, 1H, O-CH-CH₂OTs); 7.3-7.8 (m, 8H, aromatic H's).

1,5-diiodo-2(S)-ethoxy-3-oxa-pentane (C2):

TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system n-hexane/ethyl acetate (10/0.5 by volume) R_f=0.37

1HNMR (CDCl₃, 200MHz) δ:

1.24 (t, 7.0Hz, 3H, OCH₂CH₃); 3.23 (d, J=5.6Hz, 2H, J-CH₂CH-O); 3.27 (t, J=6.8Hz, 2H, J-CH₂CH₂-O); 3.58, 3.77 (two dq, J=7.0, 9.3Hz, 2H, OCH₂CH₃); 3.78, 3.87 (two dt, J=6.8, 10.6Hz, 2H, J-CH₂CH₂-O); 4.70 (t, J=5.6Hz, 1H, O-CH-CH₂-J).

Example 5

Preparation of 3'-deamino-3'[2(S)-ethoxy-4-morpholinyl] doxorubicin (A2)

Doxorubicin hydrochloride (0.5 g) was reacted with compound C2 (3 g) following the same procedure reported in Example 3 to give the title compound A2 (0.28 g) as hydrochloride salt. TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system methylene chloride/methanol (10/1 by volume) R_f=0.58

FD-MS: m/e 694 (M+)

1HNMR as free base (CDCl₃, 200MHz) δ:

1.16 (t,J=7.0Hz, 3H, OCH₂CH₃); 1.36 (d, J=6.4Hz, 3H, 5'-CH₃); 1.7-1.8 (m, 2H, 2'-CH₂); 2.16 (dd, J=4.1, 14.7Hz, 1H, 8ax-H); 2.3-2.6 (m, 6H, CH₂-N-CH₂);, 8eq-H, 3'-H); 3.02 (d, J=18.8Hz, 1H, 10ax-H); 3.26 (dd, 1.9, 18.8Hz, 1H, 10eq-H); 3.46, 3.78 (two dg, J=7.0, 9.8Hz, 2H, OCH₂CH₃); 3.53, 3.93 (two m, 2H, OCH₂CH₂N); 3.68 (s, 1H, 4'-H); 3.93 (q, J=6.4Hz, 1H, 5'-H); 4.08 (s, 3H, OCH₃); 4.56 (dd, J=2.3, 4.7Hz, 1H, OCH(OCH₂CH₃)CH₂N); 4.72 (s, 1H, 9-OH); 4.74 (s, 2H, COCH₂OH); 5.30 (m, 1H, 7-H); 5.55 (m, 1H, 1'-H); 7.3-8.1 (m, 3H, aromatic H's); 13.25 (s, 1H, 11-OH); 13.97 (s, 1H, 6-OH)

Example 6

Preparation of 1 , 5-diiodo-2(R)-isopropyloxy-3-oxa-pentane (C3)

The title compound C3 was prepared starting from L-arabinose

(3 g) following sequential reactions as described in Example

1 and Example 2.

1-isopropyl-ß-L-arabinopyranoside (S3): [α]_D= +225° (water) l,5-dihydroxy-2(R)-isopropyloxγ-3-oxa-pentane (E3):

TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system methylene chloride/methanol (10/1 by volume) R_f=0.36

1,5-di(p-toluensulphonyl)oxy-2(R)-isopropyloxy-3-oxa-pentane (F3

TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system methylene chloride/acetone (98/2 by volume) R_f=0.55

¹HNMR (CDCl₃, 200MHz) 6:

1.05, 1.10 (two d, J=6.2Hz, 6H, CH(CH₃)₂); 2.42 two s, 6H, CH₃-TS); 3.64 (two m, 2H, Ts-O-CH₂CH₂-O); 3.76 (m, 1H,

CH(CH₃)₂); 3.84 (m, 2H, O-CH-CH₂-OTs); 4.08 (t, J=5.6Hz, 2H,

Ts-O-CH₂CH₂-O); 7.3-7.8 (m, 8H, aromatic H's).

1,5-diiodo-2(R)-isopropyloxy-3-oxa-pentane (C3) TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system n-hexane/ethyl acetate (10/0.5 by volume) R_f=0.40

¹HNMR (CDCl₃, 200MHz) δ:

1.20, 1.22 (two d, J=6.4Hz, 6H, CH(CH₃)₂); 3.24 (d, J=5.6Hz,

2H, O-CH-CH₂-J); 3.28 (t, J=6.7Hz, 2H, J-CH₂CH₂-O); 3.6-3.8

(m, 2H, J-CH₂CH₂-O); 3.94 (m, 1H, CH(CH₃)₂); 4.76 (t, J=5.6Hz,

1H, O-CH-CH₂-J).

Example 7

Preparation of 3 '-deamino-3 ' [2 (R)-isopropyloxyl-4-morpholinyl] doxorubicin (A3)

Doxorubicin hydrochloride (0.5 g) was reacted with compound C3 (3.2 g) following the same procedure reported in Example

3 to give the title compound A3 (0.21 g) as hydrochloride salt.

TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system methylene chloride/methanol (10/1 by volume) R_f=0.55

FD-MS: m/e 708 (M+)

¹HNMR as free base (CDCl₃, 200MHz) δ:

1.09, 1.16 (two d, J=6.0Hz, 6H, CH(CH₃)₂); 1.36 (d, J=6.6Hz,

3H, 5'-CH₃); 1.80 (m, 2H, 2'-CH₂); 2.15 (dd, J=4.0, 14.9Hz,

1H, 8ax-H); 2.3-2.8 (m, 6H, CH₂NCH₂, 8eq-H, 3'-H); 2.97 (d,

J=18.8Hz, 1H, 10ax-H); 3.26 (d, J=18.8Hz, 1H, 10eq-H); 3.54

(m, 1H, O-CH(H)CH₂N); 3.74 (s, 1H, 4'-H); 3.81-4.1 (m, 3H,

OCH(H)CH₂N, 5'-H, OCH(CH₃)₂); 4.08 (s, 3H, OCH₃); 4.66 (s,

1H, 9-OH); 4.68 (dd, J=2.2, 4.9Hz, 1H, OCH[OCH(CH₃)₂]CH₂N);

4.75 (s, 2H, COCH₂OH), 5.28 (m, 1H, 7-H); 5.55 (m, 1H,

1'-H); 7.3-7.8 (m, 3H, aromatic H's); 13.24 (s, 1H, 11-OH);

13.97 (s, 1H, 6-OH). Example 8

1,5-dihvdroxy-2(S)-methoxy-3-oxa-pentane (E4)

1,5-dioxo-2(S)-methoxy-3-oxa-pentane (D4 ) (1.5 g, 11mmole), prepared as described in "Methods on Carbohydrate Chemistry" Acad.Press., Vol.1, 445, (1962), was dissolved in a mixture of water (10 ml) and methanol (10 ml) and treated with, sodiumborohydride (0.1 g) dissolved in water (2 ml). After 20 minutes the solution was brought to pH 7 with an acidic resin Dowex 50WX2, filtered off and the solvent was removed under reduced pressure to give 1.4 g (Yield 93%) of the title compound. TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system methylene chloride:methanol (10:1 by volume) brown spot at R_f=0.24 after heating the TLC plate previously sprayed with sulforic acid.

1HNMR (200 MHz, DMSO-d₃) δ:

3.26 (s, 3H, OCH₃); 3.4-3.6 (m, 6H, -CH₂-CH₂-O-CH-CH₂-); 4.37 (t, J=5.4 HZ, 1H, O-CH-O); 4.40 (bm, 2H, HO-CH₂CH₂, CH₂CH₂-OH)

Example 9

1,5-di(p-tolucnsulfonyl)oxy-2(S)-methoxy-3-oxa-pcntane (F4 )

1,5-dihydroxy-2(S)-methoxy-3-oxa-pentane (E4) (1.4 g, 10.3 mmole), prepared as described in Example 8, was dissolved in dry pyridine (10 ml) and treated at 0°c with p-toluensul- fonylchloride (6.4 g, 0.034 mole). The mixture was kept at 4°C overnight, then was poured into an ice-water mixture and finally extracted with methylene chloride. The organic psase was washed with water, separated off, dried over anhydrous sodium sulphate, filtered off. The solvent was removed under reduced pressure. The crude material was ehromatographed on a silicic acid column using methylene chloride as eluting agent to give 2.8 g (yield 62%) of pure titlc derivative. TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system methylene chloride: acetone (95:5 by volume) brown spot at

R_f=0.55 after heating the TLC plate previously sprayed with sulforic acid.

¹HNMR (200 MHz, CDCl₃ ) δ:

2.44 (s, 6H, CH₃-Ph); 3.27 (s, 3H, OCH₃); 3.69 (m, 2H,

SO₂OCH₂CH₂-O); 3.90 (m, 2H, SO_aOCH₂-CH-O); 4.11 (m, 2H,

SO_aOCH₂CH₂-O); 4.56 (t, J=5.3HZ, 1H, -O-CH-CH₂); 7.3-7.8 (m,

8H, aromatic H's).

Example 10

Preparation of 1 , 5-diiodo-2(s)-methoxy-3-oxa-pentane (C4 )

1,5-di(p-toluensulfonyl)oxy-2(S)-methoxy-3-oxa-pcntanc (F4 ) (1.6 g, 3.6 mmole), prepared as described in Example 9 , was dissolved in methylethylketone (30 ml) and treated with sodium iodide (3.04 g, 20.2 mmole) at 85°C for two days.

After that, the solvent was removed in vacuo and the residue was added with n-hexane (50 ml) and water. The organic phase was separated off, dried over anhydrous sodium sulphate, filtered off. The solvent was removed under reduced pressure to give 1,5-diiodo-2(S)-methoxy-3-oxa-pentane. (C42) (1.2 g, yield 86%). TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system methylene chloride, brown spot at R_f=0.54 after heating the TLC plate previously sprayed with sulforic acid. ¹HNMR (200 MHz, CDCl₃) δ:

3.15 (m, 4H, J-CH₂CH₂-OCH-CH₂-J); 3.40 (s, 3H, OCH₃); 3.80 (m, 2H, J-CH₂CH₂-O); 4.62 (t, J=5.6 Hz, 1H, O-CH-CH₂).

Example 11

Preparation of

3'-deamino-[2"(S)-methoxy-4"-morpholinyl]-doxorubicin (A4)

To a solution of doxorubicin hydrochloride (80 mg, 0.138 mmole) dissolved in dry dimethylformamide (4 ml) was added 1,5-diiodo-2(S)-methoxy-3-oxa-pentane ( C4;, 0.8 g, 2.06 mmole) and triethylamine (0.056 ml, 0.388 mmole). The reaction mixture was kept at room temperature under stirring for 36 hrs, then was poured in water and extracted with methylene chloride. After standard work up, the crude product was purified on silicic acid column using as eluting system amixture of methylene chloride methanol (97.5 : 2.5 by volume), to give, after treatment with methanolic anhydrous hydrogen chloride, 40 mg (yield 45%) of the title compound as hydrochloride salt. TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system methylene chloride:methanol (19:1 by volume) R_f=0.15

Free base: ¹HKMR (400 MHz, CDCl₃), δ:

13.98 (s, 1H, 6-OH); 13.27 (s, 1H, 11-OH); 8.03 (dd, J=1.1, 7.7Hz, 1H, 1-H), 7.78 (dd, J=7.7, 8.6Hz, 1H, 2-H); 7.39 (dd, J=1.1, 8.6Hz, 1H, 3-H); 5.55 (m, 1H, 1'-H); 5.30 (dd, J=2.1, 4.1Hz, 1H, 7-H); 4.74 (d, J=4.5Hz, 14-CH₂OH); 4.74 (s, 1H, 9-OH); 4.49 [dd, J=2.6, 4.1Hz, 1H, NCH₂-CH(OCH₃)O); 4.08 (s, 3H, 4-OCH₃); 3.94 (q, J=6.6Hz, 1H, 5'-H); 3.93 [m, 1H, NCH₂CH(H)O]; 3.67 (m, 1H, 4'-H); 3.54 [m, 1H, NCH₂CH(H)O]; 3.38 [s, 3H, NCH₂CH-OCH₃]; 3.27 (dd, J=19, 18.8Hz, 1H, 10-Heq); 3.04 (d, J=18.8Hz, 1H, 10-Hax); 3.00 (t, J=4.5Hz,1H, CH₂OH); 2.60 [dd, J=4.1, 11.4Hz, 1H, NCH(H)CHOCH₃); 2.6-2.5 (m, 3H, NCH(H)CHOCH₃, NCH₂CH₂O); 2.4-2;3 (m, 2H, 8-Heq, 3'-H); 2.15 (dd, J=4.1, 14.7Hz, 8-Hax); 1.76 (m, 2H, 2'-CH₂), 1.26 (d, J=6.6H2, 3H, 5'-CH₃).

Example 12

Preparation of 1 ,5-dihydroxy-2(R)-methoxy-3-oxa-pentane (E5)

The title compound was prepared as described in Example 8 starting from 1,5-dioxo-2(R)-methoxy-3-oxa-ρcntane (D5 ) which in turn can be prepared as described in "Methods on Carbohydrate Chemistry" Acad.Press., Vol.1, 445, (1962). TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system

methylene chloride:methanol, brown spot at R_f=0.24 after heating the TLC plate previously sprayed with sulforic acid. Example 13

Preparation of 1,5-dl(ρ-toluensulfonyl)oxy-2(R)-methoxy-

3-oxa-penfcane (F5 )

1,5-dihydroxy-2(R)-methoxy-3-oxa-pentane (E5), prepared as described in Example 12, was transformed into the title compound F5 following the same procedure reported in Example 9. TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system methylene chloride: acetone (95:5 by volume) brown spot at R_f=0.55 after heating the TLC plate previously sprayed with sulforic acid.

Example 14

Preparation of 1,5-diiodo-2(R)-methoχy-3-oxa-pentane ( C5 )

1,5-di(p-toluensulfonyl)oxy-2(R)-methoxy-3-oxa-pcntanc ( F5), prepared as described in Example 13, was converted into the diiodo derivative C5 following the procedure described in Example 10. TLC on Kieselgel Plate F₂₅₄ (Merck), eluting methylene chloride, system brown spot at R_f=0.54 after heating the TLC plate previously sprayed with sulforic acid.

Example 15

Preparation of

3'-deamino-[2"(R)-methoxy-4"-morpholinyl]-doxorubicin (A5)

The title compound was prepared by condensing doxorubicin hydrochloride with 1,5-diiodo-2(R)-methoxy-3-oxa-pcntane (C5) , prepared as above described, following the procedure reported in Example 11.

TLC on Kieselgel Plate F₂₅₄ (Merck), eluting system

methylene chloride:methanol (19:1 by volume) R_f=0.13 Free base: ¹HNMR (400 MHz, CDCl₃) δ:

13.97 (s, 1H, 6-OH); 13.25 (s, 1H, 11-OH); 8.03 (dd, J=1.1, 7.7Hz, 1H, 1-H); 7.78 (dd, J=7.6, 7.7Hz, 1H, 2-H); 7.39 (dd, J=l.l, 7.6Hz, 1H, 3-H); 5.53 (d, J=3.4Hz, 1H, 1'-H); 5.29 (dd, J=2.5, 4.1HZ, 1H, 7-H); 4.75 (s, 2H, 14-CH₂OH); 4.71 (S, 1H, 9-OH); 4.46 (dd, J-2.6, 4.7Hz,1H, NCH₂-CH(OCH₃)O]; 4.08 (s, 3H, 4-OCH₃); 3.93 (q, J=6.6Hz, 1H, 5'-H); 3.92 [m, 1H, NCH₂CH(H)O]; 3.70 (m, 1H, 4'-H); 3.56 [m, 1H, NCH₂CH(H)O); 3.40 [s, 3H, NCH_aCH-OCH₃); 3.26 (dd, J=19, 19.9Hz, 1H, 10-Heq); 3.03 (d, J=19.9Hz, 1H, 10-Hax); 2.66 [dd, J=2.6, 11.4Hz, 1H, NCH(H)CHOCH₃]; 2.53 (m, 1H, NCH(H)CH₂O); 2.5-2.3 (m, 4H, NCH(H)CH₂O, NCH(H)CHOCH₃, 8-Heq, 3'-H); 2.15 (dd, J=4.1, 14.7Hz, 8-Hax); 1.8-1.7 (m, 2H, 2'-CH₂); 1.36 (d, J=6.6Hz, 3H, 5'-CH₃).

BIOLOGICAL ACTIVITY OF 3^,-deamino-[2"(S)-methoxy-4"- morpholinyl]doxorubicin (A4) and 3'-deamino-[2"(R)- methoxy-4"-morpholinyl]doxorubicin (A5).

The compounds have been tested in several experimental system in order to ascertain their cytotoxicity and antitumor activity in experimental animals in comparison with parent Doxorubicin. The new anthracyclincs result more cytotoxic than the parent drug on LoVo and LoVo Doxorubicin- resistant cell line (LoVo/Dx), Table 1, and are active "in vivo" against doxorubicin-resistant cell lines.

The primary screening "in vivo" was carried out in BDFl mice bearing P388 Doxorubicin-resistant Johnson's leukemia (P388/Dx) (10" cell/mouse). The drugs were administered iv on day 1 after tumor inoculation. Results are reported in Table 2. Both compounds were active and more potent than Doxorubicin.

Compound A4 has also been tested on Doxorubicin-resistant P388 Schabel "in vitro". Table 3, and "in vivo" on BDFl mice (10" cell/mouse), treatment iv on day 1 after the tumor inoculation, Table 4. Finally, compound la has been studied on solid tumor such as mammary murine and human carcinoma (MX1) with iv and oral route. Table 5 and 6.

Claims

1. An anthracycline glycoside of general formula

in which X represents a linear or branched C₁-C₆ alkyl group or a benzyl group and which has a (S) or (R)

configuration at the 2"-carbon atom, and pharmaceutically acceptable acid addition salts thereof.

2. A compound according to claim 1, wherein X is methyl.

3. A compound according to claim 1, wherein X is C₂-C₆ alkyl.

4. A compound according to claim 1, which is selected from the group consisting of 3'-deamino-3'-[2"(S)- benzyloxy-4"-morpholinyl]-doxorubicin and its hydrochloride; 3'-deamino-3'-[2"(S)-ethoxy-4"-morpholinyl]-doxorubicin and its hydrochloride; 3'-deamino-3'-[2"(R)-isopropyloxy-4"- morpholinyl]-doxorubicin and its hydrochloride; 3'-deamino- 3^,-[2"(S)-methoxy-4"-morpholinyl]-doxorubicin and its hydrochloride; and 3'-deamino-3'-[2"(R)-methoxy-4"- morpholinyl] and its hydrochloride.

5. A process for preparing an anthracycline glycoside of formula A as defined in claim 1, or a

pharmaceutically acceptable salt thereof, which process comprises:

(i) reacting doxorubicin or an acid addition salt thereof with a diiodo compound of formula C:

wherein X is as defined in claim 1; and

6. A process according to claim 5, wherein doxorubicin or its hydrochloride, dissolved in a polar aprotic solvent is reacted, at room temperature and in the presence of a dry organic base, with the diiodo compound of general formula C to give the corresponding morpholinyl doxorubicin derivative of formula A which, after

purification on a silica gel column using as eluting system methylene chloride-methanol (97:5 v/v), is isolated, by treatment with methanolic anhydrous hydrogen chloride, as its hydrochloride.

7. A pharmaceutical composition comprising a pharmaceutically acceptable carrier or diluent and, as active ingredient, an anthracycline glycoside of formula A as defined in claim 1 or a pharmaceutically acceptable acid addition salt thereof.

8. An anthracycline glycoside of formula A as defined in claim 1, or a pharmaceutically acceptable acid addition salt thereof, for use as an antitumor agent.

9. A diiodo compound of formula C as defined in claim 5.

10. A process for the preparation of a diiodo compound of formula C as defined in claim 5, which process comprises:

(a) subjecting to periodate oxidation a compound of formula S¹:

wherein X is as defined in claim 1;

(b) reducing the thus-obtained dialdehyde derivative of formula D¹:

wherein X is as defined above;

(c) sulfonating the thus-obtained dihydroxy derivative of formula E¹:

wherein X is as defined above; and

(d) iodinating the sulfonated derivative thus obtained.

11. A process according to claim 10, wherein D- or L-arabinose is reacted with an alcohol X-OH, wherein X is as defined in claim 1, thereby to form the compound of formula si.

12. A process according to claim 10, wherein the dihydroxy derivative of formula E¹ is reacted with ptoluenesulfonyl chloride in step (c).